CN104017042B - A kind of separation purification method of 7-DHC - Google Patents
A kind of separation purification method of 7-DHC Download PDFInfo
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- CN104017042B CN104017042B CN201410210759.2A CN201410210759A CN104017042B CN 104017042 B CN104017042 B CN 104017042B CN 201410210759 A CN201410210759 A CN 201410210759A CN 104017042 B CN104017042 B CN 104017042B
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Abstract
The invention discloses a kind of separation purification method of 7-DHC.First be dissolved in solvent hexane and water by Biosynthesis of cholesterol gained 7-DHC fermented material, filter after fully stirring, stratification after filtering, obtains aqueous phase and organic phase; To organic phase distillation removing normal hexane, then add methyl alcohol heated and stirred, suction filtration after stirring, obtains filtrate and filter residue; To filtrate decrease temperature crystalline, suction filtration, obtain cholesterol; To add the mixture heated and stirred of normal hexane and methyl alcohol in filter residue, and carry out underpressure distillation after fully stirring, remaining material decrease temperature crystalline, suction filtration after distillation, finally obtain high purity product 7-DHC.Utilize the present invention can effectively from product 7-DHC fermented material separating-purifying go out high purity 7-DHC, and can the unconverted cholesterol of efficient recovery, make it be reused for again to feed intake, thus its raw material is utilized greatly, meet the demand of current sustainable economic development.
Description
Technical field
The present invention relates to a kind of organic method of purification, particularly relate to a kind of separation purification method of 7-DHC.
Background technology
7-DHC (7-DHC) is a kind of steroid substance.Be present in sebiferous gland in animal skin and secretory product thereof.7-DHC is white crystalline powder or light yellow crystalline powder, and its molecular formula is C
27h
44o, also known as vitamins D
3former.Can be transformed by cholesterol in human body.It is stored in subcutaneous, can change vitamins D under daylight or uviolizing
3, there is the biological activity regulating Ca,P metabolism.Therefore, children's get sun more or carry out uviolizing according to plan can preventing child rickets.
7-DHC main application is: it is synthesis of vitamin d
3important intermediate, also as the additive in skin care sun care preparations.
At present, the preparation method of 7-DHC is generally: one is that 7-DHC is by separation and Extraction in pigskin; Two is utilize cholesterol to obtain through acidylate, oxidation, hydrazone, de-hydrazone, saponification, a kind of medicine intermediate of 7-DHC saponification reaction synthesis.Utilizing cholesterol bio-transformation to synthesize 7-DHC is the synthetic method succeeded in developing at present, but does not carry out large-scale industrial production.
But utilize cholesterol to carry out in bio-transformation synthetic method, the product obtained is 7-DHC fermented material, containing microzyme culture medium, unconverted raw material cholesterol and 7-DHC in the 7-DHC fermented material obtained.Therefore, want to obtain the higher 7-DHC product of purity, need to study a kind of can the method for effective separating-purifying 7-DHC from product 7-DHC fermented material.
Summary of the invention
The technical problem to be solved in the present invention is: provide a kind of can the method for effective separating-purifying 7-DHC from product 7-DHC fermented material, namely the invention provides a kind of separation purification method of 7-DHC.Utilize technical solution of the present invention can effectively from product 7-DHC fermented material separating-purifying go out 7-DHC, and can the unconverted cholesterol of efficient recovery, make it be reused for and again feed intake.The 7-DHC product utilizing technical solution of the present invention can obtain purity to be greater than 98%.
In order to solve the problem, the technical scheme that the present invention takes is:
The invention provides a kind of separation purification method of 7-DHC, described separation purification method comprises the following steps:
A, utilize existing cholesterol bio-transformation synthetic method to obtain 7-DHC fermented material in containing microzyme culture medium, unconverted raw material cholesterol and 7-DHC; First be dissolved in solvent hexane by gained 7-DHC fermented material, and add water, the add-on of described normal hexane and water is 5 ~ 10 times of 7-DHC fermented material volume, stirs 1 ~ 1.5h at normal temperatures after adding normal hexane and water; Filter after stirring, remove water insoluble and impurity that is normal hexane; Gained filtrate stratification, then carries out separatory, separates aqueous phase and organic phase; Adopt n-hexane gained aqueous phase 2 ~ 3 times after separatory, merge organic phase;
B, the organic phase obtained by step a are distilled, and removing solvent hexane, adds the methyl alcohol of 48 ~ 52 times of volumes, be heated to 64 ~ 66 DEG C, stir 1.5 ~ 2 hours with this understanding, carry out suction filtration after stirring, obtain filtrate and filter residue after suction filtration in the product obtained;
C, the filtrate obtained by step b are cooled to room temperature and carry out crystallization, carry out suction filtration, obtain cholesterol after crystallization;
D, the filter residue obtained by step b add in the mixture of normal hexane and methyl alcohol, and the mixture volume ratio therebetween of described filter residue and normal hexane and methyl alcohol is 1:10 ~ 12, volume ratio 1:1 therebetween when described normal hexane and methanol mixed; Then be warming up to 40 DEG C ~ 45 DEG C, stir 1 ~ 1.5 hour under this temperature condition; After stirring, gained liquid distills under vacuum tightness 0.09MPa, 10 ~ 15 DEG C of conditions, is distilled to 30 ~ 35% of primary liquid volume; Then decrease temperature crystalline, carries out suction filtration after crystallization, finally obtains high purity product 7-DHC.
According to the separation purification method of above-mentioned 7-DHC, carry out suction filtration after stirring described in step b, during its suction filtration, vacuum tightness is 0.09MPa, temperature is 15 ~ 20 DEG C.
According to the separation purification method of above-mentioned 7-DHC, the crystallization time described in step c during crystallization is 12 ~ 14h.
According to the separation purification method of above-mentioned 7-DHC, described in step c, during suction filtration, vacuum tightness is 0.09MPa, temperature is normal temperature.
According to the separation purification method of above-mentioned 7-DHC, the mixture volume ratio therebetween of filter residue described in steps d and normal hexane and methyl alcohol is 1:10.
According to the separation purification method of above-mentioned 7-DHC, decrease temperature crystalline described in steps d carries out crystallization 5 ~ 6 hours for being cooled to 0 DEG C; Carry out suction filtration after described crystallization, during suction filtration, vacuum tightness is 0.09MPa, temperature is 0 DEG C.
According to the separation purification method of above-mentioned 7-DHC, the purity of the 7-DHC of high purity product described in steps d is greater than 98%.
positive beneficial effect of the present invention:
1, utilize technical solution of the present invention can effectively from product 7-DHC fermented material separating-purifying go out 7-DHC, and can the unconverted cholesterol of efficient recovery, make it be reused for again to feed intake, thus its raw material is utilized greatly, meet the demand of current sustainable economic development.
2, the 7-DHC product utilizing technical solution of the present invention can obtain purity to be greater than 98%, meets the requirement that market is higher to its product purity better.
3, adopt chemical method to produce in 7-DHC technological process at present and will use 8 kinds of poisonous and hazardous organic solvents, and the raw material of separating-purifying of the present invention adopts biotransformation method synthesis, and only use a small amount of (being generally two kinds) organic solvent in the product separation stage in biotransformation method building-up process.Decrease pollutant emission, protect physical environment, meet biological fermentation production trend in the world.
4, the present invention has all made atlas analysis to the 7-DHC standard substance that market is sold and products obtained therefrom of the present invention, and by analysis: standard substance purity is 92%, product purity of the present invention is that 98.24%(refers to accompanying drawing 1,2).
accompanying drawing illustrates:
The liquid-phase chromatographic analysis figure of Figure 17-dehydrocholesterol standard substance;
The liquid-phase chromatographic analysis figure of Fig. 2 product 7-DHC of the present invention.
embodiment:
Set forth the present invention further below in conjunction with embodiment, but do not limit content of the present invention.
Embodiment 1:
The separation purification method of 7-DHC of the present invention, the detailed step of this separation purification method is as follows:
A, utilize existing cholesterol bio-transformation synthetic method to obtain 7-DHC fermented material in containing microzyme culture medium, unconverted raw material cholesterol and 7-DHC; First be dissolved in solvent hexane by gained 7-DHC fermented material, and add water, the add-on of described normal hexane and water is 8 times of 7-DHC fermented material volume, stirs 1.2h at normal temperatures after adding normal hexane and water; Filter after stirring, remove water insoluble and impurity that is normal hexane; Gained filtrate stratification, then carries out separatory, separates aqueous phase and organic phase; Adopt n-hexane gained aqueous phase 3 times after separatory, merge organic phase;
B, the organic phase obtained by step a are distilled, removing solvent hexane, the methyl alcohol of 50 times of volumes is added in the product obtained, be heated to 65 DEG C, stir 1.8 hours with this understanding, carry out suction filtration after stirring, during suction filtration, vacuum tightness is 0.09MPa, temperature is 20 DEG C, obtains filtrate and filter residue after suction filtration;
C, the filtrate obtained by step b are cooled to room temperature and carry out crystallization 12h, carry out suction filtration after crystallization, and during suction filtration, vacuum tightness is 0.09MPa, temperature is normal temperature, obtains cholesterol;
D, the filter residue obtained by step b add in the mixture of normal hexane and methyl alcohol, and the mixture volume ratio therebetween of described filter residue and normal hexane and methyl alcohol is 1:10, volume ratio 1:1 therebetween when described normal hexane and methanol mixed; Then be warming up to 45 DEG C, stir 1 hour under this temperature condition; After stirring, gained liquid distills under vacuum tightness 0.09MPa, 15 DEG C of conditions, is distilled to 33% of primary liquid volume; Then be cooled to 0 DEG C and carry out crystallization 5 hours, carry out suction filtration after crystallization, during suction filtration, vacuum tightness is 0.09MPa, temperature is 0 DEG C, finally obtains high purity product 7-DHC (products obtained therefrom purity is 98.24%, refers to accompanying drawing 2).
Embodiment 2:
The separation purification method of 7-DHC of the present invention, the detailed step of this separation purification method is as follows:
A, utilize existing cholesterol bio-transformation synthetic method to obtain 7-DHC fermented material in containing microzyme culture medium, unconverted raw material cholesterol and 7-DHC; First be dissolved in solvent hexane by gained 7-DHC fermented material, and add water, the add-on of described normal hexane and water is 10 times of 7-DHC fermented material volume, stirs 1h at normal temperatures after adding normal hexane and water; Filter after stirring, remove water insoluble and impurity that is normal hexane; Gained filtrate stratification, then carries out separatory, separates aqueous phase and organic phase; Adopt n-hexane gained aqueous phase 2 times after separatory, merge organic phase;
B, the organic phase obtained by step a are distilled, removing solvent hexane, the methyl alcohol of 48 times of volumes is added in the product obtained, be heated to 64 DEG C, stir 2.0 hours with this understanding, carry out suction filtration after stirring, during suction filtration, vacuum tightness is 0.09MPa, temperature is 18 DEG C, obtains filtrate and filter residue after suction filtration;
C, the filtrate obtained by step b are cooled to room temperature and carry out crystallization 13h, carry out suction filtration after crystallization, and during suction filtration, vacuum tightness is 0.09MPa, temperature is normal temperature, obtains cholesterol;
D, the filter residue obtained by step b add in the mixture of normal hexane and methyl alcohol, and the mixture volume ratio therebetween of described filter residue and normal hexane and methyl alcohol is 1:11, volume ratio 1:1 therebetween when described normal hexane and methanol mixed; Then be warming up to 40 DEG C, stir 1.5 hours under this temperature condition; After stirring, gained liquid distills under vacuum tightness 0.09MPa, 10 DEG C of conditions, is distilled to 30% of primary liquid volume; Then be cooled to 0 DEG C and carry out crystallization 6 hours, carry out suction filtration after crystallization, during suction filtration, vacuum tightness is 0.09MPa, temperature is 0 DEG C, finally obtains high purity product 7-DHC.
Embodiment 3:
The separation purification method of 7-DHC of the present invention, the detailed step of this separation purification method is as follows:
A, utilize existing cholesterol bio-transformation synthetic method to obtain 7-DHC fermented material in containing microzyme culture medium, unconverted raw material cholesterol and 7-DHC; First be dissolved in solvent hexane by gained 7-DHC fermented material, and add water, the add-on of described normal hexane and water is 5 times of 7-DHC fermented material volume, stirs 1.5h at normal temperatures after adding normal hexane and water; Filter after stirring, remove water insoluble and impurity that is normal hexane; Gained filtrate stratification, then carries out separatory, separates aqueous phase and organic phase; Adopt n-hexane gained aqueous phase 3 times after separatory, merge organic phase;
B, the organic phase obtained by step a are distilled, removing solvent hexane, the methyl alcohol of 50 times of volumes is added in the product obtained, be heated to 66 DEG C, stir 1.5 hours with this understanding, carry out suction filtration after stirring, during suction filtration, vacuum tightness is 0.09MPa, temperature is 15 DEG C, obtains filtrate and filter residue after suction filtration;
C, the filtrate obtained by step b are cooled to room temperature and carry out crystallization 14h, carry out suction filtration after crystallization, and during suction filtration, vacuum tightness is 0.09MPa, temperature is normal temperature, obtains cholesterol;
D, the filter residue obtained by step b add in the mixture of normal hexane and methyl alcohol, and the mixture volume ratio therebetween of described filter residue and normal hexane and methyl alcohol is 1:12, volume ratio 1:1 therebetween when described normal hexane and methanol mixed; Then be warming up to 43 DEG C, stir 1.2 hours under this temperature condition; After stirring, gained liquid distills under vacuum tightness 0.09MPa, 15 DEG C of conditions, is distilled to 35% of primary liquid volume; Then be cooled to 0 DEG C and carry out crystallization 5 hours, carry out suction filtration after crystallization, during suction filtration, vacuum tightness is 0.09MPa, temperature is 0 DEG C, finally obtains high purity product 7-DHC.
Claims (4)
1. a separation purification method for 7-DHC, is characterized in that, described separation purification method comprises the following steps:
A, utilize existing cholesterol bio-transformation synthetic method to obtain 7-DHC fermented material in containing microzyme culture medium, unconverted raw material cholesterol and 7-DHC; First be dissolved in solvent hexane by gained 7-DHC fermented material, and add water, the add-on of described normal hexane and water is 5 ~ 10 times of 7-DHC fermented material volume, stirs 1 ~ 1.5h at normal temperatures after adding normal hexane and water; Filter after stirring, remove water insoluble and impurity that is normal hexane; Gained filtrate stratification, then carries out separatory, separates aqueous phase and organic phase; Adopt n-hexane gained aqueous phase 2 ~ 3 times after separatory, merge organic phase;
B, the organic phase obtained by step a are distilled, and removing solvent hexane, adds the methyl alcohol of 48 ~ 52 times of volumes in the product obtained, be heated to 64 ~ 66 DEG C, stir 1.5 ~ 2 hours with this understanding, carry out suction filtration after stirring, during suction filtration, vacuum tightness is 0.09MPa, temperature is 15 ~ 20 DEG C; Filtrate and filter residue is obtained after suction filtration;
C, the filtrate obtained by step b are cooled to room temperature and carry out crystallization, carry out suction filtration after crystallization, and during suction filtration, vacuum tightness is 0.09MPa, temperature is normal temperature, obtains cholesterol;
D, the filter residue obtained by step b add in the mixture of normal hexane and methyl alcohol, and the mixture volume ratio therebetween of described filter residue and normal hexane and methyl alcohol is 1:10 ~ 12, volume ratio 1:1 therebetween when described normal hexane and methanol mixed; Then be warming up to 40 DEG C ~ 45 DEG C, stir 1 ~ 1.5 hour under this temperature condition; After stirring, gained liquid distills under vacuum tightness 0.09MPa, 10 ~ 15 DEG C of conditions, is distilled to 30 ~ 35% of primary liquid volume; Then decrease temperature crystalline, carries out suction filtration after crystallization, finally obtains high purity product 7-DHC;
Described decrease temperature crystalline carries out crystallization 5 ~ 6 hours for being cooled to 0 DEG C; Carry out suction filtration after described crystallization, during suction filtration, vacuum tightness is 0.09MPa, temperature is 0 DEG C.
2. the separation purification method of 7-DHC according to claim 1, is characterized in that: the crystallization time described in step c during crystallization is 12 ~ 14h.
3. the separation purification method of 7-DHC according to claim 1, is characterized in that: the mixture volume ratio therebetween of filter residue described in steps d and normal hexane and methyl alcohol is 1:10.
4. the separation purification method of 7-DHC according to claim 1, is characterized in that: the purity of the 7-DHC of high purity product described in steps d is greater than 98%.
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| CN113015739A (en) * | 2018-11-19 | 2021-06-22 | 帝斯曼知识产权资产管理有限公司 | 7-dehydrocholesterol-hemimethanolic compounds |
| EP4139324A1 (en) * | 2020-04-23 | 2023-03-01 | DSM IP Assets B.V. | Organic solvent nanofiltration of 7-dehydrocholesterol or 25-hydroxy-7-dehydrocholesterol or their oh protected forms |
| CN112979738B (en) * | 2021-03-02 | 2022-04-05 | 浙江新和成股份有限公司 | Crystallization purification method of 7-dehydrocholesterol and application thereof in VD3 production |
| CN115746075B (en) * | 2022-10-14 | 2024-07-09 | 浙江新和成股份有限公司 | Method for recovering and purifying 7-dehydrocholesterol in photochemical reaction liquid with high impurity content in vitamin D3 production process |
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| CN102702295A (en) * | 2012-05-24 | 2012-10-03 | 河南利伟生物药业股份有限公司 | Preparation method of 7-dehydrocholesterol |
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| CN102702295A (en) * | 2012-05-24 | 2012-10-03 | 河南利伟生物药业股份有限公司 | Preparation method of 7-dehydrocholesterol |
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| 《维生素D-3合成新工艺的初步设想》;韦昌金等;《化工进展》;20031231;第22卷(第10期);第1098-1101页 * |
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Denomination of invention: A Method for Separation and Purification of 7-Dehydrocholesterol Effective date of registration: 20221109 Granted publication date: 20160106 Pledgee: Wenxian Rural Credit Cooperative Association Renmin Road Credit Cooperative Pledgor: Henan Liwei Biological Pharmaceutical Co.,Ltd. Registration number: Y2022980021218 |
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